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二级脉冲场凝胶电泳:一种更快分离更大DNA分子的新方法。

Secondary pulsed field gel electrophoresis: a new method for faster separation of larger DNA molecules.

作者信息

Zhang T Y, Smith C L, Cantor C R

机构信息

Human Genome Center, Lawrence Berkeley Laboratory, University of California, Berkeley 94720.

出版信息

Nucleic Acids Res. 1991 Mar 25;19(6):1291-6. doi: 10.1093/nar/19.6.1291.

DOI:10.1093/nar/19.6.1291
PMID:2030945
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC333856/
Abstract

A novel technique, which we call secondary pulsed field gel electrophoresis (SPFG) has been developed. In SPFG, short pulses are applied in the direction of net migration of the DNA in addition to the reorienting pulses used in conventional pulsed field electrophoresis (PFG). Experimental results show that SPFG extends and improves the electrophoretic resolution of DNA for molecules from 0.5 megabase pairs to over 10 megabase pairs in size. This improved resolution is obtained with dramatically shorter run times. Thus SPFG appears to circumvent a number of the key limitations in previous PFG protocols.

摘要

我们开发了一种名为二次脉冲场凝胶电泳(SPFG)的新技术。在SPFG中,除了传统脉冲场电泳(PFG)中使用的重新定向脉冲外,还会在DNA净迁移方向上施加短脉冲。实验结果表明,对于大小从0.5兆碱基对到超过10兆碱基对的分子,SPFG扩展并提高了DNA的电泳分辨率。这种提高的分辨率是在显著更短的运行时间内获得的。因此,SPFG似乎规避了先前PFG方案中的一些关键限制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3793/333856/b98c3558c6df/nar00242-0120-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3793/333856/4b3405918d56/nar00242-0119-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3793/333856/4788eb55324e/nar00242-0119-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3793/333856/fb845bc73034/nar00242-0119-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3793/333856/b98c3558c6df/nar00242-0120-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3793/333856/4b3405918d56/nar00242-0119-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3793/333856/4788eb55324e/nar00242-0119-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3793/333856/fb845bc73034/nar00242-0119-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3793/333856/b98c3558c6df/nar00242-0120-a.jpg

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引用本文的文献

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本文引用的文献

1
New techniques for purifying large DNAs and studying their properties and packaging.纯化大型DNA并研究其性质和包装的新技术。
Cold Spring Harb Symp Quant Biol. 1983;47 Pt 1:189-95. doi: 10.1101/sqb.1983.047.01.024.
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Optimized conditions for pulsed field gel electrophoretic separations of DNA.用于DNA脉冲场凝胶电泳分离的优化条件。
Nucleic Acids Res. 1988 Aug 11;16(15):7563-82. doi: 10.1093/nar/16.15.7563.
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Nucleic Acids Res. 1987 Jun 11;15(11):4481-9. doi: 10.1093/nar/15.11.4481.
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High-resolution separation and accurate size determination in pulsed-field gel electrophoresis of DNA. 2. Effect of pulse time and electric field strength and implications for models of the separation process.DNA脉冲场凝胶电泳中的高分辨率分离与准确大小测定。2. 脉冲时间和电场强度的影响及其对分离过程模型的启示。
Biochemistry. 1988 Dec 27;27(26):9210-6. doi: 10.1021/bi00426a020.
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High-resolution separation and accurate size determination in pulsed-field gel electrophoresis of DNA. 1. DNA size standards and the effect of agarose and temperature.DNA脉冲场凝胶电泳中的高分辨率分离及精确大小测定。1. DNA大小标准物以及琼脂糖和温度的影响。
Biochemistry. 1988 Dec 27;27(26):9204-10. doi: 10.1021/bi00426a019.
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High-resolution separation and accurate size determination in pulsed-field gel electrophoresis of DNA. 3. Effect of electrical field shape.DNA脉冲场凝胶电泳中的高分辨率分离与精确大小测定。3. 电场形状的影响。
Biochemistry. 1988 Dec 27;27(26):9216-21. doi: 10.1021/bi00426a021.
8
An electrophoretic karyotype of Neurospora crassa.粗糙脉孢菌的电泳核型。
Mol Cell Biol. 1988 Apr;8(4):1469-73. doi: 10.1128/mcb.8.4.1469-1473.1988.
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Observation of individual DNA molecules undergoing gel electrophoresis.对进行凝胶电泳的单个DNA分子的观察。
Science. 1989 Jan 13;243(4888):203-6. doi: 10.1126/science.2911733.
10
Construction of a Not I restriction map of the fission yeast Schizosaccharomyces pombe genome.粟酒裂殖酵母基因组的Not I限制酶切图谱构建
Nucleic Acids Res. 1989 Apr 11;17(7):2801-18. doi: 10.1093/nar/17.7.2801.